4.8 Article

Selection of aptamers against Ara h 1 protein for FO-SPR biosensing of peanut allergens in food matrices

Journal

BIOSENSORS & BIOELECTRONICS
Volume 43, Issue -, Pages 245-251

Publisher

ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.12.022

Keywords

Aptamer; Peanut allergen; Ara h 1; Capillary electrophoresis (CE); Systematic evolution of ligands by exponential enrichment (SELEX); Fiber optic surface plasmon resonance (FO-SPR)

Funding

  1. Flemish Institute for the Promotion of Innovation through Science and Development (Sensors for Food VIS-traject, Flanders' Food)
  2. EFRO
  3. KU Leuven (IOF-Knowledge platform Nanodiag)

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The rising prevalence to food allergies in the past two decades, together with the fact that the only existing therapy is avoidance of allergen-containing food next to the implementation of anti-allergic drugs, urges the need for improved performance of current assays to detect potential allergens in food products. Therein, the focus has been on aptamer-based biosensors in recent years. In this paper we report for the first time the selection of aptamers against one of the most important peanut allergens, Ara h 1. Several Ara h1 DNA aptamers were selected after eight selection rounds using capillary electrophoresis (CE)-SELEX. The selected aptamers specifically recognized Ara h 1 and did not significantly bind with other proteins, including another peanut allergen Ara h 2. The dissociation constant of a best performing aptamer was in the nanomolar range as determined independently by three different approaches, which are surface plasmon resonance, fluorescence anisotropy, and capillary electrophoresis (353 +/- 82 nM, 419 +/- 63 nM, and 450 +/- 60 nM, respectively). Furthermore, the selected aptamer was used for bioassay development on a home-built fiber optic surface plasmon resonance (FO-SPR) biosensor platform for detecting Ara h 1 protein in both buffer and food matrix samples demonstrating its real potential for the development of novel, more accurate aptamer-based biosensors. In conclusion, the reported aptamer holds a great potential for the detection of Ara h 1 in both the medical field and the food sector due to its high affinity and specificity for the target protein. (c) 2012 Elsevier B.V. All rights reserved.

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